Genome-wide RNAi screening identifies host restriction factors critical for in vivo AAV transduction

TY - JOUR

T1 - Genome-wide RNAi screening identifies host restriction factors critical for in vivo AAV transduction

AU - Mano, Miguel

AU - Ippodrino, Rudy

AU - Zentilin, Lorena

AU - Zacchigna, Serena

AU - Giacca, Mauro

PY - 2015/9/8

Y1 - 2015/9/8

N2 - Viral vectors based on the adeno-associated virus (AAV) hold great promise for in vivo gene transfer; several unknowns, however, still limit the vectors' broader and more efficient application. Here, we report the results of a high-throughput, whole-genome siRNA screening aimed at identifying cellular factors regulating AAV transduction. We identified 1,483 genes affecting vector efficiency more than 4-fold and up to 50-fold, either negatively or positively. Most of these factors have not previously been associated to AAV infection. The most effective siRNAs were independent from the virus serotype or analyzed cell type and were equally evident for single-stranded and self-complementary AAV vectors. A common characteristic of the most effective siRNAs was the induction of cellular DNA damage and activation of a cell cycle checkpoint. This information can be exploited for the development of more efficient AAV-based gene delivery procedures. Administration of the most effective siRNAs identified by the screening to the liver significantly improved in vivo AAV transduction efficiency.

AB - Viral vectors based on the adeno-associated virus (AAV) hold great promise for in vivo gene transfer; several unknowns, however, still limit the vectors' broader and more efficient application. Here, we report the results of a high-throughput, whole-genome siRNA screening aimed at identifying cellular factors regulating AAV transduction. We identified 1,483 genes affecting vector efficiency more than 4-fold and up to 50-fold, either negatively or positively. Most of these factors have not previously been associated to AAV infection. The most effective siRNAs were independent from the virus serotype or analyzed cell type and were equally evident for single-stranded and self-complementary AAV vectors. A common characteristic of the most effective siRNAs was the induction of cellular DNA damage and activation of a cell cycle checkpoint. This information can be exploited for the development of more efficient AAV-based gene delivery procedures. Administration of the most effective siRNAs identified by the screening to the liver significantly improved in vivo AAV transduction efficiency.

KW - Adeno-associated virus

KW - DNA-damage response

KW - High-throughput screening

KW - RNA interference

KW - Self-complementary vectors

UR - http://www.scopus.com/inward/record.url?scp=84941249203&partnerID=8YFLogxK

U2 - 10.1073/pnas.1503607112

DO - 10.1073/pnas.1503607112

M3 - Article

C2 - 26305933

AN - SCOPUS:84941249203

SN - 0027-8424

VL - 112

SP - 11276

EP - 11281

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 36

ER -